Correlation Factor Analysis Of The Symptom Of Female Gonococcal Infections

PartⅠStudy on Classification of the gonococcal genetic island ofdifferent genotypes and the relationship of GGI with femalegonococcal asymptomatic infectionsObjective Two molecular epidemiology methods which was Opa Genotyping andNG-MAST Genotyping were carried out to identify the genomic species of Neisseriagonorrhoeae.To evaluate effectivity of two molecular epidemiology methods and study therelationship between sex partner and different genotypes and female gonococcalasymptomatic infections.Methods The methods of Opa Genotyping and NG-MAST Genotyping was used todifferentiate 12 strains from male and 81 strains from female patients who attended theoutpatients of sexually transmitted disease clinics and the relationship between differentgenotypes and phenotypes was studied. Furthermore, the genetic diversity and phylogeneticrelationship of the 12 paired strains was investigated using the Clustaix software, whichwas based on the gene sequences information.Results 12 paired strains of Neisseria gonorrhoeae were classified into 10 ST genotypes byusing NG-MAST genotyping,whereas the specimens were classified into 12 OT Opagenotyping genotypes by using Opa genotyping; a new-found ST genotype (217-679)was aspecific epidemic strain in domestic. Some 80.43% of the 81 strains contained the island,different isolates from female asymptomatic infections carried different gonococcal geneticislands(positive and negative),so were the isolates from symptomatic infections.Conclusion Genotypes of each strains from patient-sex partner besides 45/46 are the samewhich indicated that contagious infection exist between patient and sex partner.Strainswithin sexual networks could be differentiated by Opa genotyping and NG-MASTgenotyping analysis. General agreement was found between these two techniques.Opa genotyping was more effective than NG-MAST genotyping in identify the genomic speciesof Neisseria gonorrhoeae.ST genotype could be further classfied into different opa-types.Different isolates carried different gonococcal genetic islands(Pathogenicity island) andcertain phenotypes, and there were little relationship between GGI and symptomaticgonococcal infections. PartⅡStudy of the relationship between the symptom of femalegonococcal infections, Chlamydia trachomatis infections and sexhormoneObjective To study the relationship between the symptom of female gonococcalinfections, Chlamydia trachomatis infections and sex hormone.Methods We studied 136 gonorrhea patients with less symptomatic (asymptomatic group)and 45 with symptomatic(symptomatic group ). Their endocervical specimens were testedfor Chlamydia trachomatis, Neisseria gonorrhoeae by PCR, the levels of TNF-αandIL-1βby ELISA. Serum progesterone (P) and estradiol(E2) levels were measured by radioimmunoassay (RIA). The results were evaluated by SPSS for Windows (version 12.0) usingstatistical analysis.Results Chlamydia trachomatis have no relationgnship with female asymptomaticgonococcal infections. However, the relationship between the decreased levels of TNF-α,IL-1βand the increased levels of progesterone was considered statistical significant at P＜0.05.Conclusion Progesterone may play a key role in female less symptomatic gonococcalinfections by down-regulatd inflammatory response. PartⅢEffect and mechanism of progesterone on PMNinflammatory responses to wild-type gonococci in vitroObjective We used 3 types of strains associated with female symptomatic Infections and 3with asymptomatic Infections to investigate the modulation effects of progesterone on PMNinflammatory responses to gonoccocus.Methods PMNs were isolated from Heparinized peripheral blood from healthy volunteers.After incubation of PMNs With 50 ng/mL 17β-estradiol for 6 hours at 37℃with 5% CO2,PR mRNA expression was determined by RT-PCR and immunofluorescence. Additionallythe effect of progesterone on N. gonorrhoeae-challenged polymorphonuclear leukocytesinflammatory responses at various time points were assayed by real-time RT-PCR. Akt,p-Akt (Ser473), iNOS and eNOS were detected respectively by Western blot.Results Among these non-unique multiple isolates from asymptomatic women(ST2951,ST735 and ST436) and from symptomatic women(ST809,ST369 and ST2948) we found nosignificant difference in Ngo-PMNs inflammatory responses. In vitro PMNs infectionexperiments, progesterone suppressed expression of iNOS,TNF-α,IL-1βtranscript andiNOS protein significantly(P＜0.01) while eNOS expression revealed a slight change. Wefound that human peripheral blood PMNs did not express the classical progesteronereceptor and the role of progesterone was independent of Akt signaling pathway.Conclusions Gonococci are susceptible to progesterone or their downstream effects invitro. Progesterone induced down-regulated expression of pro-inflammatory cytokines suchas iNOS etc., which is independent of akt signaling pathway in Ngo challanged PMNs. PartⅣEffect of progesterone on gonococci-induced apoptosisand respiratory burst of human polymorphonuclear leucocytesin vitroObjective To investigate the regulatory effect of progesterone on apoptosis and oxidativeburst activity of polymorphonuclear leucocytes(PMNs) challenged by Neisseriagonorrhoeae (Ngo) in vitro.Methods Isolated PMNs were incubated with progesterone or staurosporine. Staurosporinewas used as a positive control for our vitro model. Expression levels of inhibitory apoptosisproteins (IAPs), cellular IAP2(cIAP2) and X-linked IAP (XIAP) were determined byreal-time PCR. Additionally, PMNs apoptosis at various time points (3, 8, 12 and 24h) wereassayed by flow cytometry. Luminol amplified chemiluminescence methods were used toquantify the oxidative burst function of PMNs challenged with Ngo ST2951.Results cIAP2 was up-regulated significantly in PMNs with progesterone treatment in 3hand XIAP was up-regulated slightly compared to the medium+ST2951 group, while cIAP2was down-regulated in staurosporine-challenged PMNs. Additionally we foundprogesterone delayed the onset of apoptosis activity in Ngo ST2951 challenged PMNs,notably at 12h. No statistically significant changes in PMNs oxidative burst activity wereobserved at 10 ng/mL level of progesterone. Staurosporine enhanced the production ofsuperoxide anion (respiratory burst) of human PMNs stimulated by Ngo.Conclusion Progesterone plays an important regulatory role in the interaction of PMNs andNgo. Delayed PMNs apoptosis induced by progesterone presumably acts as a mechanismfor Ngo to avoid the innate immune response and establish long-term, low-level infection inthe female reproductive tract.